Journal of Pharmacognosy and Phytochemistry 2016; 5(6): 194-198

E-ISSN: 2278-4136 P-ISSN: 2349-8234 Studies on industrially important Guttiferae and JPP 2016; 5(6): 194-198 Palmae family Received: 18-05-2016 Accepted: 19-06-2016

Kembavimath M Kotraswamy Kembavimath M Kotraswamy, Irfan N Shaikh, Rajasaheb F Ankalgi, Department of Chemistry, G.S. Shamsunnisa R Ankalgi, Imran N Shaikh and Umar Farooq Bagwan Science College, Belgaum, India

Abstract Irfan N Shaikh Department of Chemistry, The Garcinia mangostana seed oil contains 56.2% oleic acid and 6.4% linoleic acid. The palmitic SECAB Institute of Engineering (14.8%) and stearic acid (9.0%) are major components amongst the saturated acids with smaller amounts & Technology, Vijayapura, India of capric (0.9%), lauric (2.2%), myristic (6.6%) and arachidic (3.9%). Moreover, Phoenix sylvestris, cerita mistis, chrysalidocarpus lutescens, Washingtonia filifera and phoenix rupicola belong to palmae Rajasaheb F Ankalgi family and could be compared with the oils rich in lauric acid such as cinnamon and palm kernel oils (80- Essar Laboratories and Research 90% and 45-58% respectively). Centre, Hubli, India Keywords: Guttiferae, Palmae, fatty acid, industrially important Shamsunnisa R Ankalgi Essar Laboratories and Research 1. Introduction Centre, Hubli, India One of the important facts of is their diverse pool of fatty acids. The oil seeds contains Imran N Shaikh particular fatty acids with industrially important because of their characteristic properties. The Department of Chemistry, main constituent of all the oils is the fatty acids which may include saturated, monounsaturated Mahatma Gandhi PU Science and polyunsaturated fatty acid that contribute in human physiology in different ways [1]. The College, Yadgir, India seed oils containing unusual fatty acids are industrially important as they are used in the protective coatings, plastics, cosmetics, lubricants, varieties of synthetic intermediates, Umar Farooq Bagwan Department of Chemistry, stabilizers in plastic formulations. The interesting unusual fatty acids present in high SECAB Institute of Engineering concentration of certain seed oils are being exploited for the industrial utilization. These fatty & Technology, Vijayapura, India acids of unusual structures are highly important for the production of oleochemicals [2, 3]. Guttiferae are a family of about 36 genera and 1,600 species with a pantropical distribution that occurs widely in temperate regions and have been used since ancient times as folk

remedies. It is herbaceous perennial that grows in open dry stony ground and cultivated fields [4]. Extracts of the leaves, stem bark and root bark of the plant, alone or combined with other plants are widely known for the application against a number of human ailments, such as upper respiratory tract infection, dysentery diarrhea and toothache [5]. It has traditionally been used in the treatment of burns and gastrointestinal diseases [6]. Results from recent studies [7] [8] [9] [10, 11] reporting the antinociceptive , anti-inflammatory , antioxidant , antimicrobial , and cytotoxic [12] activities demonstrate the great potential of this species for use as a medicinal plant. The palm family (Palmae or ) is a conspicuous and important feature of tropical and subtropical habitats throughout the world. In general, palms are recognized instantly by the

botanist and the layman alike, despite the fact that a great diversity of morphology exists [13] among the 191 recognized genera. The current classification of the palms , comprises six subfamilies, 14 tribes and 38 subtribes, many of which are defined in the informal classification of Moore (1973). Almost all palmate qeaved palms belong to the Coryphoideae, a subfamily of 40 genera divided among three tribes and six subtribes. The subfamily includes

one pinnate-leaved genus, Phoenix, in which the leaf lamina is split to give induplicately- folded segments, as in almost all other coryphoid palms. After exhaustive survey of literature on guttiferae and palmae family it was found that there is no report entirely on this family regarding the component fatty acids. Hence, the study of this family was undertaken with a view that it might contain some unusual fatty acids. However,

no unusual fatty acids are encountered in the present work. But this will be first report on this family as well as species. Hence, the study of this family was undertaken. This will be first report on this family as well as species examined in this investigation. Correspondence Rajasaheb F Ankalgi 2. Results and Discussions Essar Laboratories and Research Centre, Hubli, India Garcinia mangostana seed oil contains 56.2% oleic acid and 6.4% linoleic acid. The palmitic (14.8%) and stearic acid (9.0%) are major components amongst the saturated acids with ~ 194 ~

Journal of Pharmacognosy and Phytochemistry

smaller amounts of capric (0.9%), lauric (2.2%), myristic oleic (26.3%) and linoleic (11.8%) acids. (6.6%) and arachidic (3.9%). The present investigation was The seed oil of Caryota urens is rich in palmitic acid (31.1%). undertaken to study the detail pattern of component fatty The other components amongst the saturated acids are lauric acids of Garcinia xanthochymus seed oil as there is only one (3.8%), myristic (4.8%) and arachidic (0.7%). The linoleic report on the fatty acid composition of this seed oil. Sabata et acid (40.1%) and oleic (16.0%) are only unsaturated acids. al., have reported 41.0% palmitic acid as the only component Livistona rotundifolia contains 10% of lauric acid and fatty acid amongst the saturated acids [14]. The oleic acid (15.2%) of palmitic acids amongst the saturated acids with (55.45%) is only the principal component amongst the small amounts of capric (0.9%), myristic (5.7%), stearic unsaturated acids with small amount of linoleic 3.5%. But in (7.1%) and behenic (2.0%). The unsaturated acids are only the species under investigation contains 33.4% of saturated oleic (33.0%), and linoleic (26.1%). Chrysalidocarpus (Areca) acids and 62.6% of unsaturated acids. The oil form this lutescene seed oil contains (41.2%) of lauric acid, (26.4%) of species contains capric (0.9%), lauric (2.2%), myristic (6.6%), myristic acid. The next major saturated acid is palmitic acid palmitic (14.8%), stearic (9.0%) and arachidic (3.9%) acids (11.8%). This oil also contains small amount of capric and amongst the saturated acids. The unsaturated acids are only stearic acids amongst the saturated acids. The unsaturated oleic (56.2%) and linoleic (6.4%). The oleic acid is present in acids are oleic (10.5%) and linoleic (7.5%). Washingtonia major proportion. The pattern of unsaturated acids is nearly filifera, contains (83.6%) of saturated acids and 16.4% of the same as reported earlier. But there is a lot of change in the unsaturated acids. The major acids are lauric (25.8%), and proportion of saturated fatty acids. However, the detail in data palmitic (38.2%) with smaller amounts of myristic (10.9%) is given in Table-1. and stearic (6.6%) acids. However, arachidic (1.0%), and Mammea longifolia, Planch and triana seeds are rich source of behonic (1.1%) acids are also present acids are only oleic vegetable oil. The kernels on extraction with petroleum ether (5.6%) and linoleic (10.8%). yielded 75% of fatty oil. This seed oil resembles the undi or Phoenix rupicola contains (51.8%) of unsaturated acids and Domba oil obtained from the seed kernels of Calophyllum (48.2%) of saturated acids. The unsaturated are oleic (41.2%) inophyllum, another tree of guttiferae family. The seed oil of and linoleic (10.6%). This seed oil contains lauric (20.8%), Mammea longifolia, contains palmitic (16.7%) and stearic myristic (12.4%), palmitic (10.9%), stearic (3.3%), arachidic (13.3%) acids as the major component of saturated acids with (0.5%) and behenic (0.3%) acids. Licuala grandis seed oil small amounts of lauric (0.4%), mayristic (1.5%) and linoleic contains (62.6%) of unsaturated acids. The unsaturated acids (33.6%) are the major unsaturated fatty acids present in the oil are only oleic (33.4%) and linoleic (29.0%). The palmitic in small amounts. The fatty oil in different seed kernels of this (27.4%) acids is only a major component amongst the family varies from 4 to 85.9%. The Iodine value ranges from saturated acids with smaller amounts of lauric (2.5%), 36.2 to 94.0. The saponification equivalent varies from 273 to myristic (2.3%) stearic (1.9%), arachidic (1.4%) and behenic 283.2. The fatty acid composition of Mammea longifolia seed (2.1%) acids. oil resembles that of undi oil would be seen from a The member of plant family are noted for their higher comparison of the fatty acids composition of Mammea percentage of lauric (16-52%) and myristic (7-51%) acids, longifolia, undi and some other seed oils of the trees of family and low percentage of linoleic acid which rarely exceeds 10 Guttiferae (Table-1). percentage [16]. However, Norice [17] has reported different Phoenix sylvestris, cerita mistis, chrysalidocarpus lutescens, pattern of fatty acid composition in three species of Washingtonia filifera and phoenix rupicola belong to palmae (palmae), which contain lauric (0.2 to 11%), family and could be compared with the oils rich in lauric acid myristic (0.5 to 17%) and larger amounts of linoleic acid such as cinnamon and palm kernel oils [15] (80-90% and 45- (18.59%). In present investigation similar pattern of fatty acid 58% respectively). The seed oil of phoenix sylvestris contains composition as reported by morice in seed fat of (45.1%) of saturated acids with lauric acid as the principal Rhopalostylis is observed in Licuala grandis, such type of component (23.8%). It also contains smaller amounts of composition has also been found in Caryota urens and myristic (11.9%), and palmitic (9.4%) acids. Oleic acid is the Livistona rotundifolia. It may be however, emphasized that major components (41.5%) and the next in order is linoleic the variation in linoleic acid reported by Morice has been acid (13.4%) amongst the unsaturated acids. Caryota mitis further confirmed in the present work. The comparison of seed oil contains 61.9% of saturated acids with lauric acid fatty acids is given in Table 2 and 3. However, the odd chain (25.8%), as the principal component. The oil also contains fatty acids which are present in negligible quantities are not myristic (10.2%), palmitic (17.7%) and stearic (8.2%) acids taken into consideration for comparison. amongst the saturated acids. The unsaturated acids are only

Table 1: Comparative of Guttiferaceae family

Present Present References 5 10 11 12 13 14 15 Present work work work Geographical India S. India S. India India India India India India India India Sources Name of the Calophyllum Calophyllum Garcinia Garcinia Garcinia Mesua Garcinia Garcinia Garcinia Mammea species inophyllum inophyllum echinocarpa indica Morella ferrea xanthochymus mangostana xanthochymus longifolia Acids Capric ------1.9 - - Lauric 2.2 ------3.2 - 1.4 Myristic 2.4 - - - 1.3 1.2 - 7.6 1.9 2.5 Palmitic 17.9 15.8 4.7 3.5 6.2 12.6 40.0 13.8 2.1 15.7 Stearic 16.3 8.7 42.7 55.4 41.5 12.2 - 8.0 40.9 11.3 Arachidic 1.8 - - - 1.3 - - 4.9 2.0 1.9

~ 195 ~ Journal of Pharmacognosy and Phytochemistry

Behenic 1.6 ------2.0 2.1 Lignocaric ------Oleic 34.2 48.7 53.6 38.4 42.6 58.1 54.5 57.2 53.1 31.6 Linoleic 24.5 22.8 - 2.7 7.1 12.9 4.5 7.4 - 34.5 Linolenic ------4.0

Table 2: Comparative study of Palmae family References (12)

New zealand Kermadec Island (Raoul) Nor-folk islands Rhopalostylis sapida Rhopalostylis cheesemanu Trace 1.1 1.1 1.1 4.5 8.4 12.0 1.2 12.3 17.5 15.6 1.5 26.6 23.6 20.6 32.0 5.3 3.4 3.8 1.2 1.2 1.2 1.3 0.9 1.1 - 1.1 0.7 1.1 - - 0.8 34.6 31.5 33.6 3.9 18.7 18.6 19.0 60.0 1.7 1.83 1.1 2.7 - - - -

Table 3: Comparative study of Palmae family

Geographical India India India India India India India India source Name of the Phoenix Caryota Caryota Livistona Chrysalidocarpus Washingtonia Phoenix Licuala species Acids Sylvestris mitis urens rotundifolia lutescens filifera rupicola grandis Capric - - - 1.9 1.8 - - - Lauric 24.8 26.8 4.8 11.0 42.2 26.8 21.8 3.5 Myristic 12.9 11.2 4.5 6.7 27.8 11.9 13.4 3.3 Palmitic 10.4 18.7 32.1 16.2 12.8 39.2 11.9 28.4 Stearic - 9.2 5.8 8.1 2.4 7.6 4.3 2.9 Arachidic - - 1.7 - - 2.0 1.5 2.4 Behenic - - - 3.0 - 2.1 1.3 3.1 Lignocaric ------Oleic 42.5 27.3 17.0 34.0 11.5 6.6 42.2 34.4 linoleic 14.4 12.8 41.1 27.1 8.5 11.8 11.6 30.0 linolenic ------others ------

3. Experimental few crystals of an electrolyte such as sodium chloride or 3.1. Extraction of seed oil sodium acetate were added during ether extraction of liberated The seeds were cleaned air dried in the shade, crushed and mixed acids. The ether extract was washed several times with extracted thoroughly with light petroleum (40-60 0C) in a distilled water till it is free from mineral acids and id dried soxhlet apparatus for 24 hours. The solvent was removed over anhydrous sodium sulphate. The mixed acids were from the extract, the last traces under reduces pressure. The obtained from the extraction by removing the solvent under residual kernels were crushed again and extracted with a fresh reduced pressure. amount of petroleum. The process was continued till no more oil was found in the petrol extract. All the petroleum extracts 3.3. Hydrogenation and Oxidation of mixed acids were mixed together, filtered, dried over anhydrous sodium About 35-40 mgs of the mixed fatty acids with 60ml of sulphate and the solvent was removed to get the oil. The oil methanol were subjected to catalytic hydrogenation with an was then stored in cold. equal amount of palladium charcoal (20)2 in a parr-low pressure hydrogenator for about at 50-60 lbs/inch [18]. The 3.2. Isolation of mixed acids free from non-saponifiable catalyst was filtered and the solvent was removed to get the matter solid hydrogenated mixed fatty acids. A mixture of methanol, A known weight of oil was hydrolyzed at room temperature benzene and sulphuric acid (20:10:1) was added to a 100ml for 24 hours, with 1N alcoholic potassium hydroxide solution. round bottom flask containing the mixed fatty acids (80- The excess of alcohol was removed by distillation under 125mg) and refluxed for one hour on water bath. The reduced pressure and it was diluted with distilled water to get methylated mixture was diluted with water, extracted with the soap solution. From the resulting soap solution petroleum ether and washed with distilled water for several unsaponifiable matter was extracted thoroughly with ether till times [19]. The petroleum ether extract was then dried over ether layer became colourless. This extraction was repeated at anhydrous sodium sulphate, filtered and the solvent was least thrice in order to make soap solution absolutely free removed. The mixed esters were then oxidized by potassium from non-saponifiable matter. The soap solution free from permanganate using acetone as a solvent. Potassium unsaponifiable matter was acidified with pure dilute permanganate (0.2gm) was added to the mixed esters (100mg) hydrochloric acid and the precipitated mixed acids were dissolved in acetone (20 ml) and acetic acid (1ml) and extracted with ether. To prevent the formation of emulsion, refluxed for 30 min. Then the solvent was removed under

~ 196 ~ Journal of Pharmacognosy and Phytochemistry

pressure, water and a little sulphuric acid were added. The reaction can be used as a colourimetric method of estimating solution was decolourised in a stream of sulphur dioxide. The the total cyclopropene constant of an oil [24]. resulting solution was extracted with petroleum ether. The extract was dried over anhydrous sodium sulphate, filtered 3.8. Reserved phase partition chromatography (RPO) and then the solvent was removed. These oxidized mixed The chromatographic studies are based on the direction of esters were hydrolysed with 0.5N alcoholic potassium gunstone and co-workers [25]. The column contains a solid hydroxide solution and the resulting free mixed acids were support such as kieselguhr made hydrophobic with chromatographed. dichlorodimethylsilane which is further coated with paraffin. Paraffin acts as the stationary phase. The columns are loaded 3.4. Iodine value and developed as described by gunstone and sykes [25] except Wijs solution was readily prepared by dissolving 7.8 gms of that they are run and preserved at room temperature. Different iodine trichloride and 8.5 gms of iodine in warm glacial acetic concentrations of aqueous acetone equilibrated with stationary acid and make solution upto 1000ml with glacial acetic acid phase containing bromothymol blue as indicator, are used as [20]. The determination was made by weighing about 100 to eluting solvents. The sample loaded on top of the column is 200 mgs of the mixed fatty acids in a small glass capsule and developed continuously and successively with solvents of was placed in 250ml glass stoppered bottle. The fat was increasing polarity. The eluate collects in siphon (2ml) and dissolved in 10ml of chloroform and 20 ml of Wijs solution passes into a specially constructed cell where it is titrated was then added by means of pipette. A blank was also run at under a stream of nitrogen with 0.01N methanolic potassium the same time. The bottles were stoppered and left in dark for hydroxide solution using an “alga” micrometer syringe as 30 minutes. At the end of the requisite time, 15ml of 10% burette. The titration curves are drawn by plotting the eluate potassium iodide solution and 100 ml of water were added. fraction against the volume of methanolic alkali required for The liberated iodine was titrated with 0.1N sodium neutralization. The position of a peak in a titration curve is thiosulphate solution using starch solution as an indicator. characteristic of an acid or group of acids in a particular solvent system and the acids present in a mixture are 3.5. Saponification value identified by comparing of their peaks with given by known The 0.5 N alcohol potash solution was prepared by dissolving acids in the same solvent system. By summing up the titres 30 gms of pure potassium hydroxide in the minimum quantity under each peak, after allowing for the small acidity due to of water and adding 1 litre of 95% ethanol [20]. After thorough the eluating solvents, the results for the chromatograms are shaking the solution was left overnight preferably at 0 0C in calculated. In this way the hydrogenated acids, mixed acids order to precipitate any potassium carbonate, and filtered and oxidised solids are chromatographed separately, the before use. Approximately 250 to 400 mgs of mixed fatty elution curves are drawn and the results calculated. acids were weighed accurately into 250 ml alkali resistant glass flask, which was provided with a reflux condenser with 3.9. Identification of di- and tri-ethenoid acids a ground glass joint. The content of the flask were gently 3.9.1. By bromination boiled for 1 hour. The condenser was washed with little water. During chromatographic analysis, the unsaturated acids were To this mixture, 1ml of phenolphthalein solution was adder isolated from the appropriate fractions and were characterized and the excess alkali was titrated while still hot against by bromination as per the method of Bibner and [26] standard 0.5N HCL or H2SO4. Muggenthaler . The unsaturated acids (500 mg) were dissolved in dry ether (30ml) and cooled to -10 oC. A solution 3.6. Refractive index of oils of bromine in dry ether (1:10) was added drop and the Abbe’s refractometer was used for measuring the refractive temperature of the mixture was maintained below -5 oC. The indices of oils. The prisms of the instrument were cleaned solution on the standing overnight at 0 oC deposited white with toluene and the spaces between them filled with clear oil crystals. The crystals were filtered off and the filtrate was free from all air bubbles [21]. Prisms were closed and the washed with an aqueous solution of sodium thiosulphate to readings were taken when the temperature remained constant remove excess of bromine. The yellowish broom viscous for at least 3 minutes. Several readings were taken for each oil residue after removal of ether was then dissolved in petroleum and the average of all the readings were recorded as the ether (60-80 oC) and allowed to stand overnight at 0oC when it refractive index. The exact temperature was also noted down. deposited crystals of bromide. The crystalline bromides were An approximate temperature correction was made by applying then characterized by their melting points and mixed melting a correction factor, F (0.00035 for temperature around 25 0C points with authentic samples. Oleic acid gave 9,10- and 0.00036 for temperature of 40 0C or more). dibromostearic acid (m.p. 28-29 oC). Linoleic acid gave 9,10,12,13-tetrabromosteric acid 9m.p. 113-114 oC) and 3.7. Estimation of cyclopropenoid acids linolenic acid gives 9,10,12,13,15,16-hexabromostearic acid The Halphen test was originally developed as an empirical (m.p. 181-182 oC) respectively. method of testing the adulteration of adulteration of various vegetables oils by cotton seed oil [22]. Though many 3.9.2. By oxidation with one percent alkaline potassium modifications of the reagent and reaction condition have been permanganate solution described, basically the method involves heating the oil with Oxidation of unsaturated acids was carried out according to 1% solution of sulphur in carbon disulphide. If cotton seed oil the method of Lapworth and Nottram [27]. About 200mg of is present a pink colour develops. The reaction is now unsaturated acids were warmed with equal weight of sodium believed to be specific for the cyclopropene ring [23] and is a hydroxide and 12 ml of water. The mixture was cooled to -5 quick and easy method of checking whether cyclopropenoid oC with vigorous shaking. The crushed ice (75gm) was added fatty acids are present in a mixture. It is possible to use the followed by potassium permanganate (1%, 16 ml). After 5 reagent as a TLC spray. Under controlled conditions the minutes the reaction mixture was decolourised by passing a

~ 197 ~ Journal of Pharmacognosy and Phytochemistry

current of sulphur dioxide followed by the addition of against Fusarium spp. Nat Prod Commun, 2006; 1:1117- concentration hydrochloric acid (6ml). A thick white 1122. flocculent precipitate was obtained. It was drained quickly 12. Conforti F, Loizzo MR, Statti AG, Menichini F. and washed with petroleum ether. The product was Cytotoxic activity of antioxidant constituents from crystallized form rectified spirits. Oleic and linoleic acids Hypericum triquetrifolium Tura, Nat Prod Res. 2007; gave 9, 10-dihydroxystearic acid and 9, 10, 12, 13- 21:42-46. tetrahydroxy stearic acids (m.p.131-132 oC and 166-167 oC) 13. Uhl NW, Dransfield J. Genera Palmarum: after ten years. respectively. In: Henderson A., Borchsenius E (eds.) Evolution and classification of palms. Mem. New York Bot. Gard. 4. Conclusion 1999; 83:245-253. In present study, investigation on guttiferae and palmae was 14. Sabata BK, Rout MK. Proc. Inst. Chemists (India). 1963; undertaken with a hope that the fatty acids of novel structure 35:193. which would be of academic interest and might have some 15. Gunstone FD. An Introduced to Chemistry and practical utility might be discovered. The results of this Biochemistry of Fatty acids and their Glycerides investigation indicate the seed oil of Garcinia mangostana Chapman and Hall, (London) (2nd edn.). 1967, 15. seed oil contains 56.2% oleic acid and 6.4% linoleic acid. The 16. Hilditch TP, Williams PN. The chemical constitution of palmitic and stearic acid are major components amongst the Natural fats, 4th Edition, chapman and Hall (London), saturated acids with smaller amounts of capric, lauric, 1964, 341. myristic and arachidic. The seed oil of Phoenix sylvestris 17. Morice IM. Seed fats of some New Zealand and contains (45.1%) of saturated acids with acid with lauric acid Australian . Phytochem, 1970; 9:1829- as the principal component (23.8%). It also contains smaller 1833. amounts of myristic (11.9%), and palmitic (9.4%) acids. Oleic 18. Vogel IA. A text book of practical organic chemistry acid is the major components and the next in order is linoleic (Longmans, green and Co., London), 1951, 996. acid amongst the unsaturated acids. 19. Kartha ARS. The glyceride structure of natural fats. I. A technique for the quantitative determination of glyceride 5. Acknowledgements types in natural fats. J Am Oil Chem Soc. 1953; 30:280- This research work is supported by the Essar Laboratories and 282. Research Centre, Hubli-580023, INDIA and Research Center, 20. Meara NL. Modern methods of plant analysis, (springer- Dept. of Applied Chemistry, SECAB Institute of Engineering verlag, Berlin), 1955; 2:332. & Technology, Vijayapur – 586101, INDIA 21. Hopkins CY. A chapter on fats and fatty oils in encyclopedia of industrial chemical analyasis, (John 6. Reference Wiley and Sons, Inc., New York), 1971; 12:472. 1. Patil V, Gislerod HR. The importance of omega-3 fatty 22. (a) G. Halphen, Adverse effects of cyclopropenoid fatty acids in diet, Current Sci. 2006; 90:908-909. acids, J. Phram. 1897; 6:390-392. 2. Ogunniyi DS. Castor oil: A vital industrial raw material, 23. Nordby HB, Heywang BW, Kircher HW, Kemmerer AR. Bioresource Technology, 2006; 97:1086-1091. Sterculic derivatives and pink egg formation. J Am Oil 3. Osman SM, Ahmad F, Ahmad I. Oilseeds and their chem Soc. 1962; 39:183-185. Utilization, Edited by Suri, R.K.; Mathur, and K.C. 24. Bailey LV, Bittman RA, Magne FC, Skau BL. Methods Rohini, Publishing House, Dehara Dun, India. 1984, 113- for the determination of cyclopropenoid fatty acids V. A 120. spectrophotometric method for cottonseed oils based 4. Davis PH. Flora of Turkey and the East Aegean Islands. upon the Halphen-test reaction. J Am Oil Chem Soc. Edinburgh University Press. Edinburgh. 1988. 1965; 42:422-424. 5. Walker AR, Sillam R. Les plantes utiles du Gabon. Paul 25. Gunstone FD, Sykes PJ. Vegetable oils. Part VIII. The Lechevalier, Paris. 1961, 6. separation of fatty acids by reversed-phase 6. Baytop T. Therapy with Medicinal Plants in Turkey. chromatography: an empirical approach and a İstanbul University Press, İstanbul. 1999. mathematical treatment. J Chem Soc. 1960, 5050-5057. 7. Apaydın S, Zeybek U, Ince I, Elgin G, Karamenderes C, 26. Bibner, Muggenthaler, Lew Kowithsch’s Chemicals Ozturk B et al. Hypericum triquetrifolium Turra. Extract Technology and analysis of oils and fats, 1913; 1(Edn. exhibits antinociceptive activity in the mouse. J 6):585 (London). Ethnopharmacol, 1999; 3:307-312. 27. Lapworth A, Mattram EN. CCXIV.—Oxidation products 8. Ozturk B, Apaydin S, Goldeli E, Ince I, Zeybek U. of oleic acid. Part I. Conversion of oleic acid into Hypericum triquetrifolium Turra extract exhibits dihydroxystearic acid and the determination of the higher antiinflammatory activity in the rat, J Ethnopharmacol, saturated acids in mixed acids from natural sources. J 2002; 80:207-209. Chem Soc. 1925, 1628. 9. Conforti F, Statti GA, Tundis R, Menichini F, Houghton P. Antioxidant activity of methanolic extract of Hypericum triquetrifolium Turra aerial part. Fitoterapia, 2002; 6:479-483. 10. Pistelli L, Bertoli A, Morelli I, Menichini F, Musmanno RA, Di Maggio T et al. Chemical and antibacterial evaluation of Hypericum triquetrifolium Turra. Phytother Res, 2005; 19:787-791. 11. Fraternalea D, Bertoli A, Giamperi L et al., Antifungal evaluation of Hypericum triquetrifolium polar extracts

~ 198 ~